Fig. 6: NE regulates neutrophils via β2-AR–β-arrestin2 signaling to inhibit the NF-κB pathway.
From: Paraventricular nucleus CRH neurons regulate acute lung injury via sympathetic nerve–neutrophil axis
a Schematic diagram of the in vitro cell experimental procedure. Created in BioRender. Liu, T. (2025) https://BioRender.com/p48gmjd. b, c BMNs were treated with NE (1 and 10 μmol/mL) and LPS (200 ng/mL) for 2 h. Neutrophils phagocytosis (b) and ROS generation (c) were measured by flow cytometry (n = 6 per group). BMNs were pretreated with Salbutamol (1 and 10 μmol/mL) and analyzed for phagocytosis (d) and ROS generation (e) by flow cytometry (n = 6 per group). f TNF concentrations in the cell culture supernatant were measured by ELISA (n = 10 per group). g Enriched KEGG pathway interaction networks from neutrophil RNA sequencing data between control mice and hM3Dq mice treated with LPS. h Western blot analysis and relative quantities of IκBα, p-IκBα, and p65, p-p65 expression in BMNs with NE or LPS treatment (n = 3 per group). i Immunofluorescence staining of p-p65 in BMNs. Scale bar: 20 μm. j Representative image and relative quantities of β-arrestin2 in BMNs following NE or LPS treatment (n = 3 per group). Significance was determined by 1-way ANOVA with Tukey’s post-hoc multiple comparisons test. All experiments were repeated three times, yielding similar results. BMNs bone marrow-derived neutrophils, LPS lipopolysaccharide, NE norepinephrine, ROS reactive oxygen species, β2-AR β2 adrenergic receptor, TNF tumor necrosis factor.
